Hybrid peptide dendrimer systems and extrahepatic delivery

Abstract

The present invention relates to compositions that can deliver therapeutic molecules such as nucleic acids to mammalian cells, and to the human and animal body and methods of preparing and using the same. The compositions comprise nanoparticles comprising a peptide dendrimer, a nucleic acid and a lipid. The compositions of the invention find utility in the field of medicine, such as for treating cancer and autoimmune diseases.

1 . A nanoparticle comprising a peptide dendrimer, a nucleic acid and a lipid, wherein the peptide dendrimer comprises at least: a core peptide sequence, a first branching residue and two first peptide motifs,
 wherein the nanoparticle is targeted to a target cell or tissue, e.g. a myeloid, lymphoid, muscle, lung cell, a CD206+ cell and/or tumour cell, and/or a myeloid, lymphoid, muscle, lung tissue and/or tumour tissue,   
       and wherein the nanoparticle is capable of transfecting the target cell in vitro at an efficiency of at least 10%, at least 12.5%, at least 15%, at least 17.5%, at least 20%, at least 25%, at least 30%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%. 
     
     
         2 . The nanoparticle of  claim 1 , wherein the nanoparticle comprises a myeloid, lymphoid, muscle, lung, CD206+ cell or tumour cell targeting motif; optionally selected from;
 a. a muscle targeting motif, optionally comprising an ASSLNIA (SEQ ID NO:1), PYDQLRH (SEQ ID NO:2), or KAMHQMQ (SEQ ID NO:3) peptide motif;   b. an integrin targeting motif, optionally comprising an RGD or ACDCRGDCFCG (SEQ ID NO:5) peptide motif;   c. a lung targeting motif, optionally comprising the peptide sequence CGFECVRQCPERC (SEQ ID NO:4);   d. a mannose receptor targeting motif, optionally a mannose sugar or maltotriose; or   e. an antibody or target-binding fragment thereof, optionally wherein the antibody specifically binds to CD3   f. a lipid linked with a bisphosphonate (BP) group.   
     
     
         3 . The nanoparticle of  claim 1 or claim 2 , wherein the lung cell is selected from an alveolar macrophage, a ciliated cell, epithelial cell, basal cell, secretory cell, club cell, alveolar cell, fibroblast, a CD206+ cell and/or an endothelial cell. 
     
     
         4 . The nanoparticle according to any one of  claims 2 or 3 , wherein the myeloid, lymphoid, muscle, lung, CD206+ cell and/or tumour cell targeting motif is bound to a negatively charged or neutrally charged polymer or lipid. 
     
     
         5 . The nanoparticle according to  claim 4 , wherein the polymer or lipid is selected from polyglutamic acid (PGA), poly(acrylic acid), alginic acid, polyethylene glycol (PEG), or cholesteryl hemisuccinate/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, a neutrally-charged zwitterionic polymer or lipid, or a glutamic acid containing peptide. 
     
     
         6 . The nanoparticle according to  claim 4 or claim 5 , wherein the polymer is a linear or dendritic PGA. 
     
     
         7 . The nanoparticle according to  claim 4 or claim 5 , wherein the polymer is a dendritic glutamic acid containing peptide. 
     
     
         8 . The nanoparticle according to any one of  claims 2 to 7 , wherein the myeloid, lymphoid, muscle, lung, CD206+ cell or tumour cell targeting motif is covalently bound to the peptide dendrimer, or wherein the nanoparticle further comprises a second peptide comprising the myeloid, lymphoid, muscle, lung or tumour cell targeting motif. 
     
     
         9 . The nanoparticle according to  claim 8 , wherein the myeloid, lymphoid or lung cell targeting motif comprises a mannose sugar, optionally wherein the mannose sugar is covalently bound to the C-terminus and/or N-terminus of the peptide dendrimer and/or the second peptide. 
     
     
         10 . The nanoparticle according to  claim 9 , wherein the mannose sugar is covalently bound to a second peptide comprising polyglutamic acid (PGA). 
     
     
         11 . The nanoparticle according to  claim 10 , wherein the second peptide comprises linear PGA comprising 2 to 10, 2 to 25, 2 to 15 or 2 to 100 glutamic acid residues, or wherein the second peptide comprises branched PGA comprising 2 to 10, 2 to 25, 2 to 15 or 2 to 100 glutamic acid residues in each branch and/or in the core sequence, optionally wherein the second peptide comprises the structure (Ac-EEEE)2KGSGGSGGSC[(S-S)-α-D-Thiomannose]. 
     
     
         12 . The nanoparticle according to  any one of the preceding claims , wherein the lipid:nucleic acid weight/weight ratio is between 0.5:1 and 40:1, between 1:1 and 30:1, or between 2:1 and 25:1, optionally wherein the lipid:nucleic acid weight/weight ratio is at least 10:1 and wherein the nanoparticle is targeted to spleen and lung, or wherein the lipid:nucleic acid weight/weight ratio is about 23:1 and the nanoparticle is targeted to lung and spleen. 
     
     
         13 . The nanoparticle according to  any one of the preceding claims , wherein the lipid comprises two lipids, three lipids, four lipids, five lipids, or more than five lipids, optionally wherein the lipids comprise DOPE, DOTMA, DODAP, DORI, DMG-PEG and/or DOTAP. 
     
     
         14 . The nanoparticle according to  claim 13 , wherein the lipid comprises DOTMA and DOPE. 
     
     
         15 . The nanoparticle according to  claim 13 or claim 14 , wherein the lipid comprises three lipid components, or four lipid components, wherein each lipid component is selected from the group comprising DOPE, DOTMA, DODAP, DORI, DMG-PEG and DOTAP. 
     
     
         16 . The nanoparticle according to  claim 13 or claim 15 , wherein the lipid comprises DODAP, DOTMA and DOPE; or wherein the lipid comprises DODAP, DOTAP, and DOPE; or wherein the lipid comprises DODAP, DORI, and DOPE; or wherein the lipid comprises DODAP, DOTMA, DOPE and DMG-PEG; or wherein the lipid comprises DODAP, DOTAP, DOPE and DMG-PEG. 
     
     
         17 . The nanoparticle according to  any one of the preceding claims , wherein the peptide dendrimer further comprises two second branching residues and four second peptide motifs, wherein one of the second branching residues is covalently bound to one of the first peptide motifs and the other second branching residue is covalently bound to the other first peptide motif, and wherein each second branching residue is covalently bound to two second peptide motifs. 
     
     
         18 . The nanoparticle according to  claim 17 , wherein the peptide dendrimer further comprises at least four third branching residues and eight third peptide motifs,
 wherein each second peptide motif is respectively covalently bound to one of the third branching residues such that each third branching residue is covalently bound to one second peptide motif, and   wherein each third branching residue is covalently bound to two third peptide motifs.   
     
     
         19 . The nanoparticle according to  any one of the preceding claims , wherein the peptide dendrimer is selected from Table 1 or Table 1B. 
     
     
         20 . The nanoparticle according to  any one of the preceding claims , wherein the peptide dendrimer has a structure selected from G1,2-RL, 3-LR; G1,2-R; G1-RL, 2-LR; G1,2-RHL; G1-LRLR; G1,2-RF, 3-HL; G1-R;  GSC G1,2-RL, 3-LR;  RHC G1,2-R;  RHC G1-RL, 2-LR;  GSC G1,2-RHL;  GSC G1-LRLR;  GSC G1,2-RF, 3-HL; or  GSC G1-R. 
     
     
         21 . The nanoparticle according to  any one of the preceding claims , wherein the nucleic acid is:
 a. a DNA, optionally an antisense oligonucleotide, a ssDNA, a dsDNA, a plasmid, a cDNA and/or a CpG molecule; or   b. an RNA, optionally an mRNA, a circRNA, an ssRNA, a dsRNA, an sgRNA, a crRNA, a tracrRNA, a lncRNA, an siRNA, an saRNA and/or a self-amplifying RNA.   
     
     
         22 . The nanoparticle according to  any one of the preceding claims , wherein the nucleic acid encodes a chimeric antigen receptor (CAR) and/or a transcription factor. 
     
     
         23 . The nanoparticle according to  claim 22 , wherein the CAR specifically binds to carcinoembryonic antigen (CEA) or CEA cell Adhersion Molecule 5 (CEACAM5) or CEA Cell Adhesion Molecule 7 (CEACAM7). 
     
     
         24 . The nanoparticle according to  claim 22 , wherein the transcription factor is selected from interferon regulatory factor 5 (IRF5), activated IRF5, inhibitor of nuclear factor kappa B kinase subunit beta (IKK2), CCAAT enhancer binding protein alpha (CEBPA). 
     
     
         25 . A nanoparticle according to any one of  claims 1 to 24  for use in medicine. 
     
     
         26 . A nanoparticle according to any one of  claims 1 to 24  for use in a method of treating cancer, an autoimmune disease, an immune cell related disease, a lung disease and/or a myopathy. 
     
     
         27 . The nanoparticle for use according to  claim 26 , wherein the cancer comprises a solid tumour. 
     
     
         28 . The nanoparticle for use according to  claim 26 or claim 27 , wherein the cancer is a lung cancer, such as NSCLC or small cell lung cancer (SCLC), liver cancer, prostate cancer, thyroid cancer, a pancreatic cancer such as pancreatic ductal adenocarcinoma (PDAC), acute myeloid lymphoma (AML), myelodysplastic syndromes (MDS), colorectal cancer such as WT KRAS CRC, a KRAS mutated metastatic tumour, a haematological tumour, an oesophageal cancer, breast cancer, prostate cancer, bladder cancer, a tumour of the GI tract, head and neck squamous cell carcinomas (HNSCC), renal cancer, myelofibrosis, a CD206+ cancer, a melanoma, prostate cancer or anal cancer. 
     
     
         29 . The nanoparticle for use according to  claim 26 , wherein the autoimmune disease is selected from multiple sclerosis (MS), Type I or Type II Diabetes, rheumatoid arthritis, Crohn's diseases, Uveitis, Inflammatory bowel disease or Systemic lupus erythematosus, or wherein the immune related disease is Gaucher's disease, a graft versus host disease, allograft rejection, acute rejection after transplant, chronic rejection after transplant, primary graft dysfunction or chronic granulamatous disease (CGD). 
     
     
         30 . The nanoparticle for use according to  claim 26 , wherein the lung disease is cystic fibrosis, asthma, tuberculosis (TB), acute lung injury (ALI), pulmonary fibrosis, such as idiopathic pulmonary fibrosis, allergic airway disease, chronic obstructive lung disease (COPD), alpha-1 antitrypsin deficiency (AATD), pulmonary arterial hypertension, pulmonary alveolar proteinosis, fibrotic lung disease, chronic lung disease or a respiratory tract infection; or wherein the muscle disease is a muscular dystrophy or a muscle wasting disease. 
     
     
         31 . A nanoparticle according to any one of  claims 1 to 24  for use as a vaccine. 
     
     
         32 . A method of producing a coated nanoparticle that is able to transfect a target cell, the method comprising mixing a solution of peptides or peptide dendrimers with a solution of preformed nanoparticles, to form the coated nanoparticle, wherein:
 a) the preformed nanoparticles have a positive surface charge and the peptide dendrimers have a net charge that is negative, or   b) the preformed nanoparticles have a negative surface charge and the peptides or peptide dendrimers have a net charge that is positive, or   c) the surface of the preformed nanoparticles is uncharged and the peptides or peptide dendrimers comprise a hydrophobic region, or   d) the preformed nanoparticles have a positive surface charge and the peptides comprise a more than 2 and less than 100 amino acid residues.   
     
     
         33 . The method according to  claim 32 (a), wherein the peptide dendrimers comprise a PGA, or a glutamic acid containing peptide comprising a glutamic acid rich domain comprising at least 4, at least 6, or at least 8 amino acid residues in total, of which at least 2 are glutamic acid, and wherein at least 20% of the amino acid residues of the glutamic acid rich domain are glutamic acid, or the method according to  claim 31 (d), wherein the peptides comprise linear PGA comprising more than 2 and less than 100 glutamic acid residues. 
     
     
         34 . The method according to  claim 32 or 33 , wherein the peptide dendrimer comprises a myeloid, lymphoid, muscle, lung, CD206+ cell, tumour cell, bone marrow, pancreas, neuronal tissue, kidney tissue, heart tissue, liver tissue, eye, joint or prostate, or to a stem cell, pancreatic cell, neuronal cell, a kidney cell, a cardiac cell, liver cell, ocular cell, synovial cell or prostate cell targeting motif; optionally selected from;
 a. a muscle targeting motif, optionally comprising an ASSLNIA (SEQ ID NO:1), PYDQLRH (SEQ ID NO:2), or KAMHQMQ (SEQ ID NO:3) peptide motif;   b. an integrin targeting motif, optionally comprising an RGD or ACDCRGDCFCG (SEQ ID NO:5) peptide motif;   c. a lung targeting motif, optionally comprising the peptide sequence CGFECVRQCPERC (SEQ ID NO:4);   d. a mannose receptor targeting motif, optionally a mannose sugar or maltotriose;   e. an antibody or target-binding fragment thereof, optionally wherein the antibody specifically binds to CD3;   f. a bone marrow or stem cell targeting motif, optionally comprising an antibody that specifically binds CD34, and/or a lipid comprising a bisphosphonate (BP) group;   g. a pancreas targeting motif, optionally comprising a glucagon-like peptide-1 (GLP-1) homolog such as Exendin-4 (SEQ ID NO:9) or a variant or binding fragment thereof;   h. a kidney targeting motif, optionally comprising (KKEEE) 3 K;   i. a neuronal tissue or cell targeting motif, optionally comprising YTIWMPENPRPGTPCDIFTNSRGKRASNGGGG (SEQ ID NO:11) or KSVRTWNEIIPSKGCLRVGGRCHPHVNGGG (SEQ ID NO:12) or a neuronal targeting peptide comprising a Phe-Arg-Trp (FRW) motif;   j. a heart targeting or cardiac cell targeting motif, optionally comprising APWHLSSQYSRT (SEQ ID NO:13);   k. a liver targeting or hepatocyte targeting motif, optionally comprising a asialoglycoprotein receptor binder such as a N-Acetylgalactosamine (Gal-NAc) sugar, or a vitamin A moiety;   l. an eye or ocular cell targeting motif, optionally comprising CARSKNKDC (SEQ ID NO:14);   m. a joint or synovial cell targeting motif, optionally comprising an antibody that specifically binds CD44, and/or a hyaluronan sugar; or   n. a prostate targeting motif, optionally comprising PKRGFQD (SEQ ID NO:15) or SNTRVAP (SEQ ID NO:16).   
     
     
         35 . A coated nanoparticle that is able to transfect a target cell, wherein the coated nanoparticle comprises a peptide at its surface, wherein:
 a) the nanoparticle is a nanoparticle according to any one of  claims 1-24 ; and/or   b) the peptide is linear PGA comprising more than 2 and less than 100 glutamic acid residues, or the peptide is branched PGA; and/or   c) the peptide is a glutamic acid containing peptide comprising a glutamic acid rich domain comprising at least 4, at least 6, or at least 8 amino acid residues in total, of which at least 2 are glutamic acid, and wherein at least 20% of the amino acid residues of the glutamic acid rich domain are glutamic acid; and/or   d) the nanoparticle is coated with a dendrimer selected from Table 1 or Table 1B; or   e) the nanoparticle is coated with a peptide selected from Table 1A, wherein the peptide is not E100.   
     
     
         36 . The coated nanoparticle according to  claim 35 (b), (c), (d) or (e), wherein the PGA and/or glutamic acid containing peptide comprises a myeloid, lymphoid, muscle, lung, CD206+ cell, tumour cell, neuronal tissue, kidney tissue, heart tissue, neuronal cell, kidney cell, or cardiac cell targeting motif; optionally selected from;
 a. a muscle targeting motif, optionally comprising an ASSLNIA (SEQ ID NO:1), PYDQLRH (SEQ ID NO:2), or KAMHQMQ (SEQ ID NO:3) peptide motif;   b. an integrin targeting motif, optionally comprising an RGD or ACDCRGDCFCG (SEQ ID NO:5) peptide motif;   c. a lung targeting motif, optionally comprising the peptide sequence CGFECVRQCPERC (SEQ ID NO:4);   d. a mannose receptor targeting motif, optionally a mannose sugar or maltotriose;   e. an antibody or target-binding fragment thereof, optionally wherein the antibody specifically binds to CD3;   f. a bone marrow or stem cell targeting motif, optionally comprising an antibody that specifically binds CD34, and/or a lipid comprising a bisphosphonate (BP) group;   g. a pancreas targeting motif, optionally comprising a glucagon-like peptide-1 (GLP-1) homolog such as Exendin-4 (SEQ ID NO:9) or a variant or binding fragment thereof;   h. a kidney targeting motif, optionally comprising (KKEEE) 3 K;   i. a neuronal tissue or cell targeting motif, optionally comprising YTIWMPENPRPGTPCDIFTNSRGKRASNGGGG (SEQ ID NO:11) or KSVRTWNEIIPSKGCLRVGGRCHPHVNGGG (SEQ ID NO:12) or a neuronal targeting peptide comprising a Phe-Arg-Trp (FRW) motif;   j. a heart targeting or cardiac cell targeting motif, optionally comprising APWHLSSQYSRT (SEQ ID NO:13);   k. a liver targeting or hepatocyte targeting motif, optionally comprising a asialoglycoprotein receptor binder such as a N-Acetylgalactosamine (Gal-NAc) sugar, or a vitamin A moiety;   l. an eye or ocular cell targeting motif, optionally comprising CARSKNKDC (SEQ ID NO:14);   m. a joint or synovial cell targeting motif, optionally comprising an antibody that specifically binds CD44, and/or a hyaluronan sugar; or   n. a prostate targeting motif, optionally comprising PKRGFQD (SEQ ID NO:15) or SNTRVAP (SEQ ID NO:16).   
     
     
         37 . A composition comprising a nanoparticle comprising first peptide dendrimer, a second peptide dendrimer, a nucleic acid and a lipid, wherein the first and second peptide dendrimers comprise at least: a core peptide sequence, a first branching residue and two first peptide motifs. 
     
     
         38 . The composition of  claim 37 , wherein the first peptide dendrimer is a first generation peptide dendrimer comprising a core peptide sequence, a first branching residue and two first peptide motifs. 
     
     
         39 . The composition according to  claim 37 or claim 38 , wherein the second peptide dendrimer further comprises at least two second branching residues and four second peptide motifs, wherein one of the second branching residues is covalently bound to one of the first peptide motifs and the other second branching residue is covalently bound to the other first peptide motif, and wherein each second branching residue is covalently bound to two second peptide motifs. 
     
     
         40 . The composition according to any one  claims 37 to 39 , wherein the second peptide dendrimer further comprises at least four third branching residues and eight third peptide motifs, wherein each second peptide motif is respectively covalently bound to one of the third branching residues such that each third branching residue is covalently bound to one second peptide motif, and wherein each third branching residue is covalently bound to two third peptide motifs. 
     
     
         41 . The composition according to any one of  claims 37 to 40 , wherein the branching residues of the first and second peptide dendrimers are independently selected from lysine, 2,4-diaminobutyric acid, ornithine, or diaminopropionic acid. 
     
     
         42 . The composition according to any one of  claims 37 to 41 , wherein the peptide motifs of the first and second peptide dendrimers independently consist of a single amino acid, dipeptide, tripeptide or tetrapeptide motifs. 
     
     
         43 . The composition according to any one of  claims 37 to 42 , wherein the first and second peptide dendrimer peptide motifs independently comprise an amino acid with a basic side chain, a non-polar side chain, an acidic side chain, and/or an uncharged polar side chain. 
     
     
         44 . The composition according to any one of  claims 37 to 43 , wherein the first and second peptide dendrimer peptide motifs independently comprise a leucine (L), an arginine (R) and/or a histidine (H) residue. 
     
     
         45 . The composition according to any one of  claims 37 to 44 , wherein the core sequence of the first and/or second peptide dendrimer comprises at least two amino acids. 
     
     
         46 . The composition according to any one of  claims 37 to 45 , wherein the core sequence of the first and/or second peptide dendrimer comprises up to 30 amino acids. 
     
     
         47 . The composition according to any one of  claims 37 to 46 , wherein the first and/or second peptide dendrimer core peptide sequence comprises the peptide sequence RHC, GSA, or GSC. 
     
     
         48 . The composition according to any one of  claims 37 to 47 , wherein the core sequence of the first and/or second peptide dendrimer comprises an ionisable amino acid such as histidine. 
     
     
         49 . The composition according to any one of  claims 37 to 48 , wherein the nanoparticle has a polydispersity index of less than 0.6, 0.55, 0.5, 0.45, 0.4, 0.39, 0.38, 0.37, 0.36, 0.35, 0.34, 0.33, 0.32, 0.31, or 0.30. 
     
     
         50 . The composition according to any one of  claims 37 to 49 , wherein the first peptide dendrimer, when used in a first reference nanoparticle consisting of the first peptide dendrimer, a nucleic acid and a lipid, the first reference nanoparticle has a PDI of between 0.05 and 0.9, between 0.06 and 0.8, between 0.07 and 0.6, between 0.08 and 0.5, between 0.09 and 0.4, between 0.1 and 0.3, between 0.12 and 0.2, or between 0.13 and 0.15 greater than the PDI of a second reference nanoparticle consisting of the second peptide a nucleic acid and a lipid. 
     
     
         51 . The composition according to any one of  claims 37 to 50 , wherein the molar ratio with respect to the nitrogen contributed by each of the first peptide dendrimer and the second peptide dendrimer is between 10:1 and 1:10, 5:1 and 1:5, 1:4 and 4:1, 1:3 and 3:1, or 1:2 and 2:1. 
     
     
         52 . The composition according to any one of  claims 37 to 51 , wherein the molar ratio with respect to the nitrogen contributed by each of the first peptide dendrimer and the second peptide dendrimer is 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, or 4:1. 
     
     
         53 . The composition according to any one of  claims 37 to 52 , wherein the first peptide dendrimer comprises a structure set forth in Table 7 and/or the second peptide dendrimer comprises a structure set forth in Table 8. 
     
     
         54 . The composition according to any one of  claims 37 to 53 , wherein the first peptide dendrimer comprises a structure G1-LRLR; G1-R; G1,2-R; G1-RLR;  GSC G1-LRLR;  RHC G1-R;  RHC G1,2-R; or  RHC G1-RLR; and/or the second peptide dendrimer comprises the structure G1,2-RL, 3-LR; G1-RL, 2-LR;  GSC G1,2-RL, 3-LR; or  RHC G1-RL, 2-LR. 
     
     
         55 . The composition according to any one of  claims 37 to 54 , wherein the nucleic acid is selected from;
 a. a DNA, optionally an antisense oligonucleotide, a ssDNA, a dsDNA, a plasmid, a CpG molecule and/or a cDNA; or   b. an RNA, optionally an mRNA, a circRNA, a ssRNA, a dsRNA, an sgRNA, a crRNA, a tracrRNA, a lncRNA, an siRNA, an saRNA and/or a self-amplifying RNA.   
     
     
         56 . The composition according to any one of  claims 37 to 55 , wherein the nucleic acid encodes a protein or peptide, optionally wherein the protein or peptide comprises a Chimeric Antigen Receptor (CAR), a transcription factor, an antigen, a hormone, a receptor, a chimeric antigen receptor, a transcription factor and/or a cytokine such as IL-2, IL-7, IL-12, IL-15, IL-21 and/or interferon. 
     
     
         57 . The composition according to any one of  claims 37 to 56 , wherein delivery of the nucleic acid to a tissue or cell is increased by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 75%, 85%, 90%, 95%, or 100% compared to delivery of the same nucleic acid to the same tissue or cell type using a composition comprising a lipid, nucleic acid and only either the first or second peptide dendrimer. 
     
     
         58 . A composition according to any one of  claims 37 to 57  for use in medicine. 
     
     
         59 . A composition according to any one of  claims 37 to 57  for use as a vaccine. 
     
     
         60 . A composition according to any one of  claims 37 to 57  for use in a method of treating cancer, an autoimmune disease and/or a lung disease. 
     
     
         61 . The composition for use according to  claim 60 , where the cancer is a solid tumour, a lung cancer, such as NSCLC or small cell lung cancer (SCLC), liver cancer, prostate cancer, thyroid cancer, a pancreatic cancer such as pancreatic ductal adenocarcinoma (PDAC), acute myeloid lymphoma (AML), myelodysplastic syndromes (MDS), colorectal cancer such as WT KRAS CRC, a KRAS mutated metastatic tumour, a haematological tumour, an oesophageal cancer, breast cancer, prostate cancer, bladder cancer, a tumour of the GI tract, head and neck squamous cell carcinomas (HNSCC), renal cancer, a CD206+ cancer, a melanoma, prostate cancer or anal cancer. 
     
     
         62 . The composition for use according to  claim 60 , where the autoimmune disease is selected from multiple sclerosis (MS), Type I or Type II Diabetes, rheumatoid arthritis, Crohn's diseases, Uveitis, or Inflammatory bowel disease or Systemic lupus erythematosus, or wherein the immune related disease is Gaucher's disease, a graft versus host disease, allograft rejection, acute rejection after transplant, chronic rejection after transplant, primary graft dysfunction or chronic granulamatous disease (CGD), or wherein the lung disease is cystic fibrosis, asthma, tuberculosis (TB), acute lung injury (ALI), pulmonary fibrosis, such as idiopathic pulmonary fibrosis, allergic airway disease, chronic obstructive lung disease (COPD), alpha-1 antitrypsin deficiency (AATD), pulmonary arterial hypertension, pulmonary alveolar proteinosis, fibrotic lung disease, chronic lung disease or a respiratory tract infection; or wherein the muscle disease is a muscular dystrophy or a muscle wasting disease. 
     
     
         63 . A nanoparticle comprising a peptide dendrimer, a nucleic acid and a lipid, wherein the peptide dendrimer comprises at least: a core peptide sequence, a first branching residue and two first peptide motifs,
 wherein the nanoparticle is targeted to bone marrow, pancreas, neuronal tissue, kidney tissue, heart tissue, liver tissue, eye, joint or prostate, or to a stem cell, pancreatic cell, neuronal cell, a kidney cell, a cardiac cell, liver cell, ocular cell, synovial cell or prostate cell.   
     
     
         64 . The nanoparticle according to  claim 63 , wherein the nanoparticle comprises a targeting motif selected from:
 a. a bone marrow or stem cell targeting motif, optionally comprising an antibody that specifically binds CD34, and/or a lipid comprising a bisphosphonate (BP) group;   b. a pancreas targeting motif, optionally comprising a glucagon-like peptide-1 (GLP-1) homolog such as Exendin-4 (SEQ ID NO:9) or a variant or binding fragment thereof;   c. a kidney targeting motif, optionally comprising (KKEEE) 3 K;   d. a neuronal tissue or cell targeting motif, optionally comprising YTIWMPENPRPGTPCDIFTNSRGKRASNGGGG (SEQ ID NO:11) or KSVRTWNEIIPSKGCLRVGGRCHPHVNGGG (SEQ ID NO:12) or a neuronal targeting peptide comprising a Phe-Arg-Trp (FRW) motif;   e. a heart targeting or cardiac cell targeting motif, optionally comprising APWHLSSQYSRT (SEQ ID NO:13);   f a liver targeting or hepatocyte targeting motif, optionally comprising a asialoglycoprotein receptor binder such as a N-Acetylgalactosamine (Gal-NAc) sugar, or a vitamin A moiety;   g an eye or ocular cell targeting motif, optionally comprising CARSKNKDC (SEQ ID NO:14);   h a joint or synovial cell targeting motif, optionally comprising an antibody that specifically binds CD44, and/or a hyaluronan sugar; or   i a prostate targeting motif, optionally comprising PKRGFQD (SEQ ID NO:15) or SNTRVAP (SEQ ID NO:16).   
     
     
         65 . The nanoparticle according to  claim 63 or claim 64 , wherein the nanoparticle comprises a cell targeting motif that targets the stem cell, pancreatic cell, neuronal cell, a kidney cell, a cardiac cell, liver cell, ocular cell, synovial cell or prostate cell. 
     
     
         66 . The nanoparticle according to any one of  claims 63 to 65 , wherein the nanoparticle is capable of transfecting the targeted stem cell, pancreatic cell, neuronal cell, a kidney cell, a cardiac cell, liver cell, ocular cell, synovial cell or prostate cell in vitro at an efficiency of at least 10%, at least 12.5%, at least 15%, at least 17.5%, at least 20%, at least 25%, at least 30%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%. 
     
     
         67 . The nanoparticle according to any one of  claims 63 to 66 , wherein the bone marrow, pancreas, neuronal tissue, kidney tissue, heart tissue, liver tissue, eye, joint or prostate, stem cell, pancreatic cell, neuronal cell, a kidney cell, a cardiac cell, liver cell, ocular cell, synovial cell or prostate cell targeting motif is bound to a negatively charged or neutrally charged polymer or lipid. 
     
     
         68 . The nanoparticle according to  claim 67 , wherein the polymer or lipid is selected from polyglutamic acid (PGA), poly(acrylic acid), alginic acid, polyethylene glycol (PEG), or cholesteryl hemisuccinate/1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, a neutrally-charged zwitterionic polymer or lipid, or a glutamic acid containing peptide. 
     
     
         69 . The nanoparticle according to  claim 67 or claim 68 , wherein the polymer is a linear or dendritic PGA. 
     
     
         70 . The nanoparticle according to  claim 67 or claim 68 , wherein the polymer is a dendritic glutamic acid containing peptide. 
     
     
         71 . The nanoparticle according to any one of  claims 63 to 70 , wherein the bone marrow, pancreas, neuronal tissue, kidney tissue, heart tissue, liver tissue, eye, joint or prostate, stem cell, pancreatic cell, neuronal cell, a kidney cell, a cardiac cell, liver cell, ocular cell, synovial cell or prostate cell targeting motif is covalently bound to the peptide dendrimer, or wherein the nanoparticle further comprises a second peptide comprising the bone marrow, pancreas, neuronal tissue, kidney tissue, heart tissue, liver tissue, eye, joint or prostate, stem cell, pancreatic cell, neuronal cell, a kidney cell, a cardiac cell, liver cell, ocular cell, synovial cell or prostate cell targeting motif. 
     
     
         72 . The nanoparticle according to  claim 71 , wherein the second peptide comprises linear PGA comprising 2 to 10, 2 to 25, 2 to 15 or 2 to 100 glutamic acid residues, or wherein the second peptide comprises branched PGA comprising 2 to 10, 2 to 25, 2 to 15 or 2 to 100 glutamic acid residues in each branch and/or in the core sequence, optionally wherein the second peptide comprises the structure (Ac-EEEE)2KGSGGSGGSC[(S-S)-α-D-Thiomannose]. 
     
     
         73 . The nanoparticle according to any one of  claims 63 to 72 , wherein the lipid:nucleic acid weight/weight ratio is between 0.5:1 and 40:1, between 1:1 and 30:1, or between 2:1 and 25:1, optionally wherein the lipid:nucleic acid weight/weight ratio is at least 10:1 and wherein the nanoparticle is targeted to spleen and lung, or wherein the lipid:nucleic acid weight/weight ratio is about 23:1 and the nanoparticle is targeted to lung and spleen. 
     
     
         74 . The nanoparticle according to any one of  claims 63 to 73 , wherein the lipid comprises two lipids, three lipids, four lipids, five lipids, or more than five lipids, optionally wherein the lipids comprise DOPE, DOTMA, DODAP, DORI, DMG-PEG and/or DOTAP. 
     
     
         75 . The nanoparticle according to  claim 74 , wherein the lipid comprises DOTMA and DOPE. 
     
     
         76 . The nanoparticle according to  claim 74 or claim 75 , wherein the lipid comprises three lipid components, or four lipid components, wherein each lipid component is selected from the group comprising DOPE, DOTMA, DODAP, DORI, DMG-PEG and DOTAP. 
     
     
         77 . The nanoparticle according to  claim 75 or claim 76 , wherein the lipid comprises DODAP, DOTMA and DOPE; or wherein the lipid comprises DODAP, DOTAP, and DOPE; or wherein the lipid comprises DODAP, DORI, and DOPE; or wherein the lipid comprises DODAP, DOTMA, DOPE and DMG-PEG; or wherein the lipid comprises DODAP, DOTAP, DOPE and DMG-PEG. 
     
     
         78 . The nanoparticle according to any one of  claims 63 to 77 , wherein the peptide dendrimer further comprises two second branching residues and four second peptide motifs, wherein one of the second branching residues is covalently bound to one of the first peptide motifs and the other second branching residue is covalently bound to the other first peptide motif, and wherein each second branching residue is covalently bound to two second peptide motifs. 
     
     
         79 . The nanoparticle according to  claim 78 , wherein the peptide dendrimer further comprises at least four third branching residues and eight third peptide motifs,
 wherein each second peptide motif is respectively covalently bound to one of the third branching residues such that each third branching residue is covalently bound to one second peptide motif, and wherein each third branching residue is covalently bound to two third peptide motifs.   
     
     
         80 . The nanoparticle according to  any one of the preceding claims , wherein the peptide dendrimer is selected from Table 1 or Table 1B, or wherein the nanoparticle is coated with a peptide selected from Table 1A. 
     
     
         81 . The nanoparticle according to any one of  claims 63 to 80 , wherein the peptide dendrimer has a structure selected from G1,2-RL, 3-LR; G1,2-R; G1-RL, 2-LR; G1,2-RHL; G1-LRLR; G1,2-RF, 3-HL; G1-R;  GSC G1,2-RL, 3-LR;  RHC G1,2-R;  RHC G1-RL, 2-LR;  GSC G1,2-RHL;  GSC G1-LRLR;  GSC G1,2-RF, 3-HL; or  GSC G1-R. 
     
     
         82 . The nanoparticle according to any one of  claims 63 to 81 , wherein the nucleic acid is:
 a. a DNA, optionally an antisense oligonucleotide, a ssDNA, a dsDNA, a plasmid, a cDNA and/or a CpG molecule; or   b. an RNA, optionally an mRNA, a circRNA, an ssRNA, a dsRNA, an sgRNA, a crRNA, a tracrRNA, a lncRNA, an siRNA, an saRNA and/or a self-amplifying RNA.   
     
     
         83 . The nanoparticle according to any one of  claims 63 to 82 , wherein the nucleic acid encodes a chimeric antigen receptor (CAR) and/or a transcription factor. 
     
     
         84 . The nanoparticle according to  claim 83 , wherein the CAR specifically binds to carcinoembryonic antigen (CEA) or CEA cell Adhersion Molecule 5 (CEACAM5) or CEA Cell Adhesion Molecule 7 (CEACAM7). 
     
     
         85 . The nanoparticle according to  claim 83 , wherein the transcription factor is selected from interferon regulatory factor 5 (IRF5), activated IRF5, inhibitor of nuclear factor kappa B kinase subunit beta (IKK2), CCAAT enhancer binding protein alpha (CEBPA). 
     
     
         86 . A nanoparticle according to any one of  claims 63 to 85  for use in medicine. 
     
     
         87 . A nanoparticle according to any one of  claims 63 to 85  for use as a vaccine. 
     
     
         88 . A nanoparticle according to any one of  claims 63 to 85  for use in a method of treating cancer, an autoimmune disease, an immune cell related disease, a lung disease and/or a myopathy. 
     
     
         89 . The nanoparticle for use according to  claim 88 , wherein the cancer comprises a solid tumour. 
     
     
         90 . The nanoparticle for use according to  claim 88 or claim 89 , wherein the cancer is a lung cancer, such as NSCLC or small cell lung cancer (SCLC), liver cancer, prostate cancer, thyroid cancer, a pancreatic cancer such as pancreatic ductal adenocarcinoma (PDAC), acute myeloid lymphoma (AML), myelodysplastic syndromes (MDS), colorectal cancer such as WT KRAS CRC, a KRAS mutated metastatic tumour, a haematological tumour, an oesophageal cancer, breast cancer, prostate cancer, bladder cancer, a tumour of the GI tract, head and neck squamous cell carcinomas (HNSCC), renal cancer, myelofibrosis, a CD206+ cancer, a melanoma, prostate cancer or anal cancer. 
     
     
         91 . The nanoparticle for use according to  claim 88 , wherein the autoimmune disease is selected from multiple sclerosis (MS), Type I or Type II Diabetes, rheumatoid arthritis, Crohn's diseases, Uveitis, or Inflammatory bowel disease, or wherein the immune related disease is Gaucher's disease, a graft versus host disease, allograft rejection, acute rejection after transplant, chronic rejection after transplant, primary graft dysfunction or chronic granulamatous disease (CGD). 
     
     
         92 . The nanoparticle for use according to  claim 88 , wherein the lung disease is cystic fibrosis, asthma, tuberculosis (TB), acute lung injury (ALI), pulmonary fibrosis, such as idiopathic pulmonary fibrosis, allergic airway disease, chronic obstructive lung disease (COPD), a Ipha-1 antitrypsin deficiency (AATD), pulmonary arterial hypertension, pulmonary alveolar proteinosis, fibrotic lung disease, chronic lung disease or a respiratory tract infection; or wherein the muscle disease is a muscular dystrophy or a muscle wasting disease.